Background <p>Our study aimed to investigate the immediate effects of a single session of Pain Neuroscience Education (PNE) on pain intensity, pain knowledge, pain catastrophizing, and kinesiophobia in undergraduate physiotherapy students with chronic non-specific low back pain (CNSLBP).</p> Methods <p>This prospective, single-arm, pre-post intervention study was conducted with 79 undergraduate physiotherapy students with CNSLBP. Participants received a single, 70-minute PNE session, administered by an expert physiotherapist with a PNE certification. Before and immediately after the intervention measurements were made using the Numeric Pain Rating Scale (NPRS), the Revised Pain Neurophysiology Questionnaire (rNPQ), the Pain Catastrophizing Scale (PCS), and the Tampa Scale of Kinesiophobia (TSK).</p> Results <p>After PNE, NPRS (<i>p</i> = 0.004; d = 0.131) and TSK (<i>p</i> = 0.034; d = 0.253) scores decreased statistically significantly, while the rNPQ score increased significantly (<i>p</i> &lt; 0.001; d = 0.505). However, there was no statistically significant improvement in the PCS score (<i>p</i> = 0.312; d = 0.124).</p> Conclusion <p>This study suggests that a single PNE session was associated with immediate improvements in pain knowledge and immediate reductions in pain intensity and kinesiophobia in undergraduate physiotherapy students with CNSLBP. Overall, PNE may be incorporated into rehabilitation as a useful educational primer to support pain-related outcomes, but it should be complemented by other interventions to achieve clinically meaningful reductions in pain intensity.</p> Trial Registration <p>ClinicalTrials.gov identifier: NCT07249710; retrospectively registered on 18 November 2025.</p>